This application is based upon and claims the benefit of priority from the prior Japanese Patent Applications No. 2000-080955, filed Mar. 22, 2000; and No. 2001-062372, filed Mar. 6, 2001, the entire contents of both of which are incorporated herein by reference.
The present invention relates to a carrier for gene detection, a method for detecting validity of interferon therapy on individuals, a gene detecting apparatus for detecting validity of interferon therapy on individuals, and a gene detection kit for detecting validity of interferon therapy.
In recent years, patients infected with hepatitis C viruses (to be called HCV hereafter) have rapidly increased, leading to a great social problem.
Although interferon therapy has been revealed to have a certain effect on hepatitis C as well as on other viral illnesses, interferon therapy is not effective to all the infected patients, and there are not a few patients who has no sensitivity to interferon and cannot benefit from interferon therapy. Continuation of interferon therapy to such patients who exhibit no sensitivity to interferon would not only cause side effects such as fever and anemia to the patients but also delay initiation of other expected therapies.
Therefore, there has been longed for a method of predicting whether interferon therapy is effective or not to an HCV-infected patient to be treated, but such prediction has not at all been possible.
An object of the invention is to provide a method for predicting whether interferon therapy is effective to a patient, before the treatment is effected.
According to the first aspect of the present invention, there is provided a carrier for gene detection, which comprises:
a base body; and
a polynucleotide immobilized on the base body, the polynucleotide comprising a polynucleotide selected from the group consisting of:
(at) the polynucleotide of Sequence ID No. 1 in the sequence listing;
(bt) a modified polynucleotide derived from the polynucleotide (at) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ct) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 1;
(dt) a polynucleotide containing the sequence which spans from 449the to 459th position of Sequence ID No. 1; and
(et) a complementary strand of the polynucleotide selected from the group consisting of (at), (bt), (ct) and (dt) mentioned above.
According to the second aspect of the present invention, there is provided a carrier for gene detection which comprises:
a base body; and
a polynucleotide immobilized on the base body, the polynucleotide comprising a polynucleotide selected from the group consisting of:
(ag) the polynucleotide of Sequence ID No. 2 in the sequence listing;
(bg) a modified polynucleotide derived from the polynucleotide (ag) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cg) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 2;
(dg) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 2; and
(eg) a complementary strand of the polynucleotide selected from the group consisting of (ag), (bg), (cg) and (dg) mentioned above.
According to the third aspect of the present invention, there is provided a carrier for gene detection which comprises:
a base body; and
a polynucleotide immobilized on the base body, the polynucleotide comprising a polynucleotide selected from the group consisting of:
(aa) the polynucleotide of Sequence ID No. 3 in the sequence listing;
(ba) a modified polynucleotide derived from the polynucleotide (aa) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ca) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 3;
(da) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 3; and
(ea) a complementary strand of the polynucleotide selected from the group consisting of (aa), (ba), (ca) and (da) mentioned above.
According to the fourth aspect of the present invention, there is provided a carrier for gene detection which comprises:
a base body; and
a polynucleotide immobilized on the base body, the polynucleotide comprising a polynucleotide selected from the group consisting of:
(ac) the polynucleotide of Sequence ID No. 4 in the sequence listing;
(bc) a modified polynucleotide derived from the polynucleotide (ac) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cc) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 4;
(dc) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 4; and
(ec) a complementary strand of the polynucleotide selected from the group consisting of (ac), (bc), (cc) and (dc) mentioned above.
According to the fifth aspect of the present invention, there is provided a DNA chip, which comprises:
a base body; and
a first and a second electrodes formed on the base body,
the first electrode comprising a conductive body and at least one polynucleotide immobilized on the conductive body, the polynucleotide being selected from the group consisting of (at) to (et) shown below;
(at) the polynucleotide of Sequence ID No. 1 in the sequence listing;
(bt) a modified polynucleotide derived from the polynucleotide (at) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ct) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 1;
(dt) a polynucleotide containing the sequence which spans from 449the to 459th position of Sequence ID No. 1; and
(et) a complementary strand of the polynucleotide selected from the group consisting of (at), (bt), (ct) and (dt) mentioned above, the second electrode comprising a conductive body, and at least one polynucleotide immobilized on the conductive body, the polynucleotide being selected from the group consisting of (ag) to (eg), (aa) to (ea), and (ac) to (ec) shown below;
(ag) the polynucleotide of Sequence ID No. 2 in the sequence listing;
(bg) a modified polynucleotide derived from the polynucleotide (ag) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cg) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 2;
(dg) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 2;
(eg) a complementary strand of the polynucleotide selected from the group consisting of (ag), (bg), (cg) and (dg) mentioned above;
(aa) the polynucleotide of Sequence ID No. 3 in the sequence listing;
(ba) a modified polynucleotide derived from the polynucleotide (aa) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ca) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 3;
(da) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 3;
(ea) a complementary strand of the polynucleotide selected from the group consisting of (aa), (ba), (ca) and (da) mentioned above;
(ac) the polynucleotide of Sequence ID No. 4 in the sequence listing;
(bc) a modified polynucleotide derived from the polynucleotide (ac) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cc) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 4;
(dc) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 4; and
(ec) a complementary strand of the polynucleotide selected from the group consisting of (ac), (bc), (cc) and (dc) mentioned above.
According to the sixth aspect of the present invention, there is provided a method for detecting validity of interferon therapy for an individual, the method comprises:
1) contacting a polynucleotide sample taken from the individual with the carrier for gene detection according to the present invention; and
2) determining the nucleotide sequence of the polynucleotide in the sample, by detecting the hybridization reaction between the polynucleotide sample and the polynucleotide immobilized on the carrier for gene detection.
Further, according to the seventh aspect of the present invention, there is provided a method for detecting validity of interferon therapy for an individual, the method comprises:
1) contacting the probe polynucleotide to a carrier for gene detection which has a polynucleotide sample taken from the individual immobilized on a substrate; and
2) determining the nucleotide sequence of the polynucleotide sample by detecting the hybridization reaction between the polynucleotide sample immobilized on the substrate and the probe polynucleotide;
The probe polynucleotide comprises a polynucleotide selected from the group consisting of:
(at) the polynucleotide of Sequence ID No. 1 in the sequence listing;
(bt) a modified polynucleotide derived from the polynucleotide (at) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ct) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 1;
(dt) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 1;
(et) a complementary strand of the polynucleotide selected from the group consisting of (at), (bt), (ct) and (dt) mentioned above;
(ag) the polynucleotide of Sequence ID No. 2 in the sequence listing;
(bg) a modified polynucleotide derived from the polynucleotide (ag) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cg) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 2;
(dg) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 2;
(eg) a complementary strand of the polynucleotide selected from the group consisting of (ag), (bg), (cg) and (dg) mentioned above;
(aa) the polynucleotide of Sequence ID No. 3 in the sequence listing;
(ba) a modified polynucleotide derived from the polynucleotide (aa) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(ca) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 3;
(da) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 3;
(ea) a complementary strand of the polynucleotide selected from the group consisting of (aa), (ba), (ca) and (da) mentioned above;
(ac) the polynucleotide of Sequence ID No. 4 in the sequence listing;
(bc) a modified polynucleotide derived from the polynucleotide (ac) by including one or several deletions, substitutions or additions at any positions except for 455th position;
(cc) a polynucleotide containing the sequence which spans from 441st to 455th position of Sequence ID No. 4;
(dc) a polynucleotide containing the sequence which spans from 449th to 459th position of Sequence ID No. 4; and
(ec) a complementary strand of the polynucleotide selected from the group consisting of (ac), (bc), (cc) and (dc) mentioned above.
According to the eighth aspect of the present invention, there is provided a gene detecting apparatus for detecting validity of interferon therapy, the apparatus comprising:
the carrier for gene detection of the present invention described above;
a reaction section for contacting a first polynucleotide immobilized on a base body of the carrier with a sample which contains a second polynucleotide labeled with a marker, and putting the first and the second polynucleotides under hybridization reaction condition; and
a marker-detecting apparatus for detecting the marker attached to the second polynucleotide.
According to the ninth aspect of the present invention, there is provided a gene detection apparatus for detecting validity of interferon therapy, the apparatus comprising:
a carrier for gene detection of the present invention described above, which is used as an electrode;
a counter electrode;
a voltage application means for applying voltage between the carrier for gene detection and the counter electrode,
a reaction section for contacting a first polynucleotide immobilized on a base body of the carrier with a sample which contains a second polynucleotide, and putting the first and the second polynucleotides under hybridization reaction condition; and
a measurement section for measuring an electric signal generated between the carrier for gene detection and the counter electrode when voltage is applied by the voltage applying means after the hybridization reaction.
According to the tenth aspect of the present invention, there is provided a kit for determining validity of interferon therapy, which comprises the carrier for gene detection described above and a buffer solution.
According to the eleventh aspect of the present invention, there is provided a kit for detecting validity of interferon therapy for an individual, which comprises the carrier for gene detection described above, a double-strand recognizer, and a buffer solution.